Team:BGU Israel/Description

>


<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">

Team:BGU Israel/Team


Team:BGU Israel




Tell us about your project, describe what moves you and why this is something important for your team.


What should this page contain?

    Overview

    This summer we have set our goal to design and test a synthetic machine which could distinguish individual cancer cells from healthy tissue, and that works only when inside the nucleus of a human cancer cells. We have constructed two separate designs: the first, for reducing tumor proliferation and inducing apoptosis by knock-down of vital genes. The second is for expression of exogenous proteins which could color the tumor in a visible way for the naked eye (for example, a chromophore), or lead to cell apoptosis by expression of an apoptotic protein. The first design utilizes CRISPR-cas9 and gRNA to knock-down a vital gene, namely, Ubb gene which encodes for poli-Ubiquitin. Ubiquiting levels are elevated in most, if not all human cancer cells, are essential to the growth of cancer cells and the protein is thought to help cancer cells adapt to increased stress. Ubb downregulation by si-RNA has shown a great decrease in tumor proliferation and more than 50 percent increase in apoptotic cells (1).

    The Design

    The design includes two parts: one with cas9 gene, and the other with a gRNA compatible to three sequences in the second axon of Ubb. Cas9 – the cas9 endonuclease was put under control of h-tert promoter, thus should be expressed predominantly in cells in which it is highly active, namely – cancer cells. When guided with gRNA, cas9 cuts both strands of DNA at the target sites, which leads to the cell trying to fix the double strand break, adding mutations which damage the activity of the mature protein. gRNA – the guide RNA is a hundred bases long molecule with a unique two dimensional structure which binds cas9 and guides it to a dsDNA sequence complementary to 22 base pairs on the 5' end of the molecule.

    The second design includes the gRNA and two different parts. dCas9-V64 – dCas9-V64 was engineered so that it lacks endonuclease activity and has 4 V16 activation domains attached. When guided to a specific promoter, dCas9-V64 promotes transcription of genes downstream of binding site. The third part is synthetic promoter regulating GFP. The synthetic promoter has 3 complementary sites for the gRNA from the second part, which, upon binding, should paint cancer cells fluorescent green. **This construct was made as a proof of concept. By utilizing a synthetic promoter we could, in theory, express an apoptotic protein, a toxin and pretty much everything with a promoter.
  • A detailed explanation of why your team chose to work on this particular project.
    • Cancer is the number one cause of mortality in developed countries
  • References and sources to document your research.
  • Use illustrations and other visual resources to explain your project.

Advice on writing your Project Description

We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.

Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.


References

(1) Downregulation of ubiquitin level via knockdown of polyubiquitin gene Ubb as potential cancer therapeutic intervention Choongseob Oh , Soonyong Park , Eun Kyung Lee2 & Yung Joon Yoo

Inspiration

See how other teams have described and presented their projects:

Retrieved from "https://2015.igem.org/Team:BGU_Israel/Team"